Fig 4. 5' and 3' rapid amplification of cDNA ends (RACE) of Rffl-lnc1.

(A) Primer design for 5’RACE and 3’RACE. P1 and P2 were designed for 5’RACE. P3 and P4 were designed for 3’RACE. (B) 5’RACE PCR amplification. Lane 1 shows the PCR products by using P1 and UPM to amplify 5’RACE cDNA. Lane 2 and 3 show the distinct 5’RACE PCR products a, b, c and d by using P2 and UPS to amplify the diluted PCR product from lane 1 (nested PCR). (C) 3’RACE PCR amplification. Lane 4 shows PCR products by using P3 and UPM to amplify 3’RACE cDNA. Lane 5 shows the distinct 3’RACE PCR product by using P4 and UPS to amplify the diluted PCR product from lane 4 (nested PCR). (D) The schematic showing four Rffl-lnc1 transcripts. The full length sequences of Rffl-lnc1 were obtained by in-fusion cloning of RACE products for sequencing.